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Regulatory statements vary by product. Please see product-specific literature to determine product use.

What positive transfection control can I use with Lipofectamine RNAiMAX? Product FAQ

Answer

BLOCK-iT Alexa Fluor Red Fluorescent Control (Cat. No. 14750100) can be used as an indicator of transfection efficiency of siRNA. This dye can be detected using standard filter sets designed for Alexa Fluor 555, Cy3, DsRed, Texas Red, or Rhodamine Red fluorophores.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Answer Id: E9041

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Can I use the PLUS Reagent with Lipofectamine RNAiMAX? Product FAQ

Answer

The PLUS Reagent will not enhance transfection efficiency when used with Lipofectamine RNAiMAX.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Answer Id: E9040

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How much Lipofectamine RNAiMAX reagent is required for a genome-wide screen? Product FAQ

Answer

It is expected that 800 plates of 96 wells would be used for a genome-wide screen. Allowing for 3 biological replicates, the total number of wells to be transfected are: 800 plates x 96 wells x 3 replicates = 230,400 wells.
0.15 µL of Lipofectamine RNAiMAX reagent needs to be added per well = 34,560 µL of Lipofectamine RNAiMAX reagent= approximately 35 mL total of Lipofectamine RNAiMAX reagent for a single genome-wide screen with 3 replicates.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

Answer Id: E10045

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Can I use Lipofectamine RNAiMAX Transfection Reagent if I accidentally froze it at -20 degrees C? Product FAQ

Answer

No, freezing may affect its performance. Lipofectamine RNAiMAX should only be stored at 4 degrees C.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Answer Id: E19756

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I accidentally froze my Lipofectamine RNAiMAX Reagent. Can I still use it? Product FAQ

Answer

Freezing can alter the integrity and composition of the lipid particles. The performance of the reagent may be affected and it may only work for the first week or so. It is safer to use a new vial.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Answer Id: E9072

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Can I use Lipofectamine RNAiMAX to co-transfect siRNA with plasmid DNA? Product FAQ

Answer

While Lipofectamine 3000 or Lipofectamine 2000 may be used for co-transfection of siRNA with plasmid DNA, Lipofectamine RNAiMAX cannot be used.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Answer Id: E9039

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I accidentally kept my Lipofectamine RNAiMAX Reagent at room temperature. Can I still use it? Product FAQ

Answer

Yes, all of our lipid transfection reagents are stable at room temperature for months.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Answer Id: E9073

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I'm seeing cell death after transfection using Lipofectamine RNAiMAX Reagent. What should I do? Product FAQ

Answer

Remove antibiotics from the medium during transfection, as this can cause stress leading to cell death. Try adjusting both lipid and siRNA quantities. Use cells with lower passage number for better viability. Consider assaying for microbes such as Mycoplasma that are not detected easily in cell culture.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Answer Id: E9074

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Can Lipofectamine Stem Transfection Reagent be used for siRNA delivery? Product FAQ

Answer

We recommend using Lipofectamine RNAiMAX Transfection Reagent for siRNA delivery.

Answer Id: E15206

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What should I use to dilute my RNAi duplexes and Lipofectamine RNAiMAX before complexing? Product FAQ

Answer

We recommend using Opti-MEM I Reduced Serum Medium (Cat. No. 31985062).

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Answer Id: E9038

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Since Silencer Select siRNA is considered more potent than Stealth siRNA, do you recommend using a lower final concentration of Silencer Select siRNA than Stealth siRNA with Lipofectamine RNAiMAX Transfection Reagent? Product FAQ

Answer

Yes, this is correct. The Lipofectamine RNAiMAX transfection protocol was written with Silencer Select siRNAs in mind, with a recommended final siRNA concentration of 5-10 nM. For other siRNAs (Silencer and Stealth siRNAs), the recommended final siRNA concentration ranges are 50-100 nM (Silencer siRNA) and 20-40 nM (Stealth siRNA). These potency (recommended working concentration) values are listed on this webpage (https://www.thermofisher.com/us/en/home/life-science/rnai/synthetic-rnai-analysis.html), under the table "Choose from our pre-designed siRNA product lines". The exact siRNA concentration to use will depend on the cell type and the particular siRNA. Some Stealth siRNAs will work at 10 nM final concentration in easy-to-transfect cell lines, but often, more will be needed.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Answer Id: E20036

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Can Lipofectamine LTX be used in RNAi experiments? Product FAQ

Answer

Lipofectamine LTX performs well with vector-based RNAi experiments since you are transfecting DNA. But for direct transfection of RNA molecules like siRNA and Stealth RNAi negative control transfections, we recommend Lipofectamine RNAiMAX.

Answer Id: E4994

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What transfection reagent should I use for transfecting siRNA? Product FAQ

Answer

The optimal transfection agent depends upon the cell type used. That being said, we find that Lipofectamine RNAiMAX transfection agent provides excellent transfection efficiency with low toxicity in most cell types.

Answer Id: E6585

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What lipid does Invitrogen recommend for RNAi applications? Product FAQ

Answer

We currently recommend Lipofectamine RNAiMAX, which is specifically developed for siRNA transfection. It provides the following advantages:
- High transfection efficiencies in many cell types to minimize background expression from untransfected cells and maximize knockdown.
- Minimal cytotoxicity to reduce non-specific effects and cellular stress.
- Generally requires low concentrations of RNAi duplexes to obtain high knockdown levels, further minimizing non-specific effects.
- A broad peak of optimal transfection activity with minimal cytotoxicity, allowing achievement of high knockdown levels despite differences in cell density, minor pipetting inaccuracies, and other variations.

Answer Id: E4174

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Can I use Lipofectamine 3000 in RNAi experiments? Product FAQ

Answer

Lipofectamine 3000 works well for delivery of vector-based RNAi and synthetic siRNA, as well as co-transfection of siRNA with plasmid DNA. However, Lipofectamine RNAiMAX is our best transfection reagent for synthetic siRNA. For in vivo siRNA experiments, we recommend Invivofectamine 3.0 Reagent.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Answer Id: E9016

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